RU2641963C2 - Cyclosporine suspensions a of form 2 - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: group of inventions represents: an ophthalmic suspension, containing cyclosporine A of form 2 and some medium, where cyclosporine A of form 2 is characterized by peaks in X-ray diffractometer scanning with the radiation of Cu Kα, 2θ: 7.5, 8.8, 10.2, 11.3, 12.7, 13.8, 14.5, 15.6 and 17.5; method of preparation of cyclosporine composition, which includes the stages of mixing cyclosporine A of form 2 with the medium for producing the suspension; suspension quassation; ophthalmic suspension, containing particles of cyclosporine A of form 2; and the medium, in which the average particle size (d90) is less than about 10 micron; method of treatment of a condition selected from: dry eye syndrome, blepharitis, tarsal glands disease, impaired awareness of corneal, allergic conjunctivitis, atopic keratoconjunctivitis, spring keratoconjunctivitis and pterygium, including the stage of applying said ophthalmic suspension to the patient, being in this condition.

EFFECT: increased bioavailability, reduced foreign body sensation and achieved high stability.

15 cl, 18 dwg, 2 ex, 2 tbl

Description

CROSS REFERENCE TO A RELATED APPLICATION

This patent application claims the priority of provisional patent application US No. 61/559 866, filed November 15, 2011, the full contents of which are incorporated herein by reference.

FIELD OF THE INVENTION

This invention relates to the fields of nanotechnology and drug technology.

BACKGROUND

Cyclosporin A is an active agent in Restasis®, a drug used to treat dry eye syndrome. Cyclosporin A is slightly soluble in water, and therefore, it is currently introduced into pharmaceutical formulations, either by dissolving the drug substance in oil to form an emulsion, or by mixing the drug substance with significant amounts of surfactants and / or solubilizers to an aqueous solution formed. The inventors invented the composition of cyclosporin A, using a new crystalline polymorph of cyclosporin A, forming nanosuspensions containing cyclosporin particles with an average particle size of about 1 micrometer or less (in order to imagine this number, the average thickness of a human hair is about 100 micrometers) .

A new suspension of cyclosporin A, when delivered topically to the eye, may have one or more advantages, including the following:

- higher bioavailability, compared with suspensions, due to the larger surface area upon dissolution;

- Longer held on the eye due to small particles, which leads to a further improvement in bioavailability;

- reduced stress when sensing a foreign body, or irritation by particles, thereby reducing lacrimation and drainage of formulations from the eye;

- a reduced content of surfactants or solubilizers in the composition, improving tolerance and bioavailability.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1 illustrates the size distribution of cyclosporin A particles of Form 2 in nanosuspensions prepared using a high pressure homogenizer, compared with suspensions of pre-grinding.

Figure 2 illustrates the particle size distribution of cyclosporin A of Form 2 in nanosized formulations prepared using various surfactants or stabilizers.

Figure 3 illustrates the particle size distribution of the Composition diluted in a medium containing Na CMC.

Figure 4 illustrates the average particle size of the cyclosporin A nanosuspension of Form 3, compared with a nanosuspension using Form 2 after micronization using a microfluidizer. Form 2 forms a nanosuspension (particle size <1 μm), while Form 3 does not.

Figures 5-13 illustrate the results when four different formulations according to the invention (summarized in Table 2) and Restasis® are used on NZW (New Zealand breed) females (two rabbits in total, one composition per eye) in a single topical dose :

Figure 5 illustrates the concentration in the cornea.

Figure 6 summarizes the pharmacokinetic data.

Figure 7 illustrates concentrations in the bulbar conjunctiva.

Figure 8 illustrates concentrations in the palpebral conjunctiva.

Figure 9 summarizes the pharmacokinetic data for bulbar conjunctiva and palpebral conjunctiva.

Figure 10 illustrates the concentration in the bulbar conjunctiva.

Figure 11 illustrates concentrations in the palpebral conjunctiva.

Figure 12 illustrates the concentration in the lacrimal gland.

Figure 13 summarizes the pharmacokinetic data for the lacrimal gland.

Figure 14 illustrates the characteristics of CsA X-ray powder diffraction (RPD) patterns in a new crystalline form (called Form 2 herein), in tetragonal form (called Form 1 in this document), and in orthorhombic form (called Form 3 in this document).

Figure 15 illustrates the RPD diffraction pattern of a CsA crystalline Form 2.

Figure 16 illustrates the characteristic of the sorption / desorption of water for CsA Form 2.

Figure 17 illustrates the analysis of MDSK CsA Form 2, extracted from 0.04% of the composition, with 1% PS80.

Figure 18 illustrates a reproduced RPD diagram formed by cyclosporin A.

DETAILED DESCRIPTION

Cyclosporin A

Cyclosporin A (CsA) is a cyclic peptide having the following chemical structure:

Its chemical name is cyclo [[(E) - (2S, 3R, 4R) -3-hydroxy-4-methyl-2- (methylamino) -6-octenoyl] -L-2-aminobutyryl-N-methylglycyl-N- methyl-L-leucyl-L-valyl-N-methyl-L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl] . It is also known under the names: “cyclosporine”, “cyclosporine A”, “ciclosporin”, and “ciclosporin A”. He is an active agent in Restasis® (Allergan, Inc., Irwin, Calif.), An emulsion containing 0.05% (w / v) cyclosporin. Restasis® is approved in the United States to increase the production of tears in patients whose tear secretion is thought to be suppressed due to ocular inflammation associated with drying keratoconjunctivitis.

Cyclosporin A Form 2

It is known that cyclosporin A exists in amorphous form, in the form of a liquid crystal, a tetragonal crystalline form (form 1), and an orthorhombic form (form 3). Recently discovered a new crystalline form - cyclosporin A form 2.

The RPD diagram of the CsA of Form 2 is significantly different from the tetragonal shape and the orthorhombic shape (FIG. 14). The main crystalline peaks for CsA form 2 appear in (2θ) when scanned by an X-ray diffractometer with an X-ray source such as Cu Kα radiation, λ = 1.54 Å, at 30 kV / 15 mA: 7.5, 8.8, 10.2, 11.3, 12.7 , 13.8, 14.5, 15.6 and 17.5 (the d-distance in the crystal lattice is approximately 11.8, 10.0, 8.7, 7.8, 7.0, 6.4, 6.1, 5.6 and 5.1 - Å, respectively, Fig. 15). These main peaks are defined as unique to Form 2, with respect to orthorhombic, or tetragonal forms; as well as peaks with an intensity exceeding the background by more than 5 times.

In one embodiment, the new crystalline form (Form 2) of CsA is a non-stoichiometric hydrate of Cyclosporin A. In another embodiment, crystalline Form 2 is represented by the following formula:

where X is the number of water molecules, ranging from 0 to 3. In one embodiment, X in the formula is 2.

Form 2 is a kinetically stable form of CsA in aqueous suspensions. Suspensions containing Form 2 do not exhibit conversion to other known polymorphic or pseudo-morphic forms during storage. It was found that Form 1 and the amorphous form are converted to Form 2 in the presence of water.

The structure of a single crystal of the hydrated form CsA of Form 2 was determined and the parameters of this crystal structure are listed in Table 2. These results indicate that Form 2 is unique compared to other known crystalline forms of Cyclosporin A.

The asymmetric CsA unit cell of Form 2 contains one cyclosporin A molecule and two water molecules. It is possible that any small molecules capable of forming a hydrogen bond with water can play the role of a volume filler, which would give a number of potential structures, ranging from orthorhombic dihydrate to distorted monoclinic dihydrate. The RPD diagram calculated from the single crystal structure is illustrated in Figure 8 and it corresponds to the experimental diagram illustrated in Figure 2. These matching diagrams further confirm that Form 2 is a unique and pure crystalline form of cyclosporin A.

Without being limited by theory, thermogravimetric analysis, combined with KF titration and analysis of steam desorption sorption (SDP), suggest that CsA Form 2 - is a non-stoichiometric hydrate and CsA. The analysis of steam sorption for Cyclosporin Form 2 shows that the water content in the new crystalline form is reversibly dependent on relative humidity, as illustrated in FIG. 16. Similarly, with the tetragonal form, this new CsA form undergoes a phase transition to a liquid crystal, or an amorphous form at 124.4 ° C before melting, as illustrated by modulated differential calorimetric analysis (MDSK) (Figure 17).

Cyclosporin Form 2 can be obtained by suspending amorphous 0.05% cyclosporin A (mass / volume) in 1% Polysorbate 80, by heating the solution to 65 ° C, keeping it at this temperature for 24 hours, and then isolating precipitate by vacuum filtration. In the future, you can use this cyclosporin A Form 2, obtained in such a way as to grow additional quantities, using Cyclosporin A Form 2 as a seed crystal; in this method, about 30 g of cyclosporin A is suspended in a solution of 900 ml of water containing 1% (mass / volume) of Polysorbate 80, this solution is heated to 65 ° C, and then 0.2 g of cyclosporin A is seeded with Form 2 at a temperature of 52 ° C. Subsequently, this solution is stirred for about 22 hours at a temperature ranging from 61 ° C to 65 ° C, and then the precipitate formed is isolated.

More details regarding CsA Form 2 can be found in US Application No. 13/480 710, the entire contents of which are incorporated herein by reference.

Suspend Cyclosporin A Form 2

The compositions of the invention are ophthalmologically acceptable formulations of Cyclosporin A Form 2. By "ophthalmologically acceptable", the inventors mean that suspensions are prepared in such a way as to be non-irritating to the eye of a mammal, such as a human. In one embodiment, pharmaceutical formulations are suspensions; that is, they are composed of cyclosporin A particles of Form 2, with an average particle size slightly larger than about 1 μm dispersed in a liquid medium. In another embodiment, the pharmaceutical compositions are nanosized; that is, they are composed of cyclosporin A particles of Form 2, with an average particle size slightly less than about 1 μm dispersed in a liquid medium. In one implementation, the suspension contains cyclosporin A form 2 with a concentration of from about 0.001% to about 10% (mass / volume). In one embodiment, the suspension contains cyclosporin A Form 2 with a concentration of from about 0.001% (mass / volume) to about 0.01%, from about 0.001% (mass / volume) to about 0.04% (mass / volume), from about 0.001% (mass / volume) to about 0.03% (mass / volume), from about 0.001% (mass / volume) to about 0.02% (mass / volume), or from about 0.001% (mass / volume) to about 0.01% (mass / volume). In another implementation, the suspension contains cyclosporin A form 2 with a concentration of from about 0.01% (mass / volume) to about 0.05%, from about 0.01% (mass / volume) to about 0.04% (mass / volume), from about 0.01% (mass / volume) to about 0.03% (mass / volume), from about 0.01% (mass / volume) to about 0.02% (mass / volume), or from about 0.01% (mass / volume) to about 0.01% (mass / volume).

In another implementation, the suspension contains cyclosporin A form 2 with a concentration of from about 0.01% (mass / volume) to about 0.1%, from about 0.1% (mass / volume) to about 0.5% (mass / volume), from about 0.01% (mass / volume) to about 1% (mass / volume), or from about 1% (mass / volume) to about 10%.

For example, the compositions may contain about 0.001% (mass / volume), about 0.002% (mass / volume), about 0.003% (mass / volume), about 0.004% (mass / volume), about 0.005% (mass / volume), about 0.006% (mass / volume), about 0.007% (mass / volume), about 0.008% (mass / volume), about 0.009% (mass / volume), about 0.01% (mass / volume), about 0.015% (mass / volume), about 0.02% (mass / volume), about 0.025% (mass / volume), about 0.03% (mass / volume), about 0.035% (mass / volume), about 0.04 % (mass / volume), about 0.045% (mass / volume), about 0.05% (mass / volume), about 0.055% (mass / volume), about 0.06% (mass / volume), about 0.065% (mass / volume), about 0.07% (mass / volume) , about 0.075% (mass / volume), about 0.08% (mass / volume), about 0.085% (mass / volume), about 0.09% (mass / volume), about 0.095% (mass / volume), about 0.1% (mass / volume), about 0.15% (mass / volume), about 0.2% (mass / volume), about 0.25% (mass / volume), 0.3% (mass / volume), about 0.35% (mass / volume), about 0.4% (mass / volume), about 0.45% (mass / volume), about 0.5% (mass / volume), about 0 , 55% (mass / volume), about 0.6% (mass / volume), about 0.65% (mass / volume), about 0.7% (mass / volume), about 0.75% (mass / volume), about 0.8% (mass / volume), about 0.85% (mass / volume), about 0.9% (mass / volume), about 0.95% (mass / volume), or about 1.0% (mass / vol ) Ciclosporin A mold 2.

In one embodiment, the suspension contains a surfactant. In one embodiment, the surfactant is selected from polyoxyethylene (20), sorbitan monooleate (Polysorbate 80), glycol polyethylene 660, hydroxystearate (Solutol), polyoxyethylene (40) Myrj 52 stearate (POE-40-Stearate), Pluronic F68 (Polaxamer 188), polyoxyethylene sorbitan monolaurate (Polysorbate 20), sodium glycocholate (NaGC). The medium may contain all of these surfactants, or one, two, three, four or five of them.

Can be used in the range of from about 0.001% (mass / volume) to about 5% (mass / volume) of a surfactant. In one embodiment, the suspension contains from about 0.001% (mass / volume) to about 1% (mass / volume), from about 0.001% (mass / volume) to about 0.1% (mass / volume), from about 0 , 01% (mass / volume) to about 0.1% (mass / volume), or from about 0.1% (mass / volume) to about 1% (mass / volume) of a surfactant. For example, suspensions may contain about 0.001% (mass / volume), about 0.002% (mass / volume), about 0.003% (mass / volume), about 0.004% (mass / volume), about 0.005% (mass / volume), about 0.006% (mass / volume), about 0.007% (mass / volume), about 0.008% (mass / volume), about 0.009% (mass / volume), about 0.01% (mass / volume), about 0, 02% (mass / volume), about 0.03% (mass / volume), about 0.04% (mass / volume), about 0.05% (mass / volume), about 0.06% (mass / volume ), about 0.07% (mass / volume), about 0.08% (mass / volume), about 0.09% (mass / volume), about 0.1% (mass / volume), about 0.2 % (mass / volume), 0.3% (mass / volume), about 0.4% (mass / volume), about 0.5% (ma csa / volume), about 0.6% (mass / volume), about 0.7% (mass / volume), about 0.8% (mass / volume), about 0.9% (mass / volume), about 1% (mass / volume), about 2% (mass / volume), about 3% (mass / volume), about 4% (mass / volume), or about 5% (mass / volume) of a surfactant.

When using more than one surfactant, the suspension may contain equal or different amounts of each of them.

In addition to surfactants, the suspension may contain a stabilizer. In one embodiment, the stabilizer is selected from hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, polyvinylpyrrolidone, carboxymethyl cellulose, Pemulen®, and Pemulen® TR-2. Pemulen® - are the trade names for high molecular weight crosslinked acrylic acid-C10-C30 alkyl acrylate copolymers manufactured by Lubrizol Corp. Pemulen® TR-2 - is a copolymer of C10-30 alkyl acrylate containing more hydrophobic groups than other Pemulen® polymers. The medium may contain all of these stabilizers, or none of them, or may contain one, two, three, four or five of them.

Between about 0.01% (w / v) and about 10% (w / v) of the stabilizer can be used. In one embodiment, the suspension contains from about 0.01% (mass / volume) to 1% (mass / volume), or from about 0.01% (mass / volume) to 0.1% (mass / volume), or

from about 0.1% (weight / volume) to about 1% (weight / volume) of the stabilizer. For example, a suspension may contain about 0.01% (mass / volume), about 0.02% (mass / volume), about 0.03% (mass / volume), about 0.04% (mass / volume), about 0.05% (mass / volume), about 0.06% (mass / volume), about 0.07% (mass / volume), about 0.08% (mass / volume), about 0.09% ( mass / volume), about 0.1% (mass / volume), about 0.2% (mass / volume), about 0.3% (mass / volume), about 0.4% (mass / volume), about 0.5% (mass / volume), about 0.6% (mass / volume), about 0.7% (mass / volume), about 0.8% (mass / volume), about 0.9% (mass / volume), about 1% (mass / volume), about 2% (mass / volume), about 3% (mass / volume), about 4% (mass / volume), about 5% (mass / volume m) about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), or about 10% stabilizer.

When using more than one surfactant, the suspension may contain equal or different amounts of each of them.

In addition to the surfactant, the medium may also contain a tonicity regulator selected from glycerol, mannitol, sodium citrate dihydrate, potassium chloride, boric acid, and sodium borate decahydrate. The tonicity regulator is added because it is necessary to achieve the desired state; a medium may contain all of these tonicity controllers, or none of them, or it may contain one, two, three, four, or five of them. In one embodiment, tonicity controls are present in an amount ranging from about 0.1% (weight / volume) to about 10% (weight / volume). When using more than one tonicity regulator, the suspension may contain equal or different amounts of each of them.

The suspension usually contains water in an amount sufficient to provide the desired pH, tonicity and other characteristics that would make the suspension suitable for use with the eye.

Methods for preparing suspensions of cyclosporin A form 2.

The compositions of this invention can be prepared by mixing cyclosporin A form 2 with the corresponding surfactants, stabilizers and tonicity regulators, as described above, to form a suspension. If the smallest particles of Cyclosporin A are required, the suspension is crushed using a high pressure homogenizer, for example, available from Microfluidics Int'l Corp. Newton, Massachusetts. The unique and amazing property of Cyclosporin A Form 2 is that it can be crushed, if necessary, to obtain a suspension with an average particle size (d90) of less than 1 μm. Cyclosporin A in such a nanosuspension has a higher bioavailability compared to other (macro) suspensions of cyclosporin A, due to the larger surface area available for dissolution; bioavailability is further improved, since small particles allow Cyclosporin A to remain in the eye longer. Smaller particles of nanosuspensions lead to the fact that the composition is less likely to cause a foreign body sensation, or other irritations perceived by the subject when the composition is instilled into the eye.

In addition, since the particles are smaller, they are better associated with surfactants and stabilizers, which allows their lower concentrations to be used.

After the suspension of cyclosporin A form 2 is crushed, it is diluted to obtain the final product.

Treatment methods

The compositions of this invention can be used to treat any condition of the eye, for which it is known that it is subject to topical treatment with cyclosporine A (for example, using Restasis®) in the concentrations mentioned here. For example, the pharmaceutical compositions of this invention can be used to treat patients suffering from dry eye syndrome, to treat blepharitis and meibomian gland disease, to restore the sensitivity of a cornea damaged due to refractive eye surgery, to treat allergic conjunctivitis, atopic keratoconjunctivitis and spring keratoconjunctivitis, for the treatment of pterygium, inflammation of the conjunctiva and cornea, keratoconjunctivitis, graft versus host, post-transplant glaucoma , corneal transplantation, fungal keratitis, Tijeson's superficial punctate keratitis, among other conditions.

The International Dry Eye Symposium (CVG) defines dry eye as "a multifactorial disease of tears and the ocular surface, manifested by symptoms of discomfort, visual impairment, and instability of the tear film with potential damage to the ocular surface, accompanied by increased osmosis of the tear film and inflammation of the ocular surface." It includes conditions such as dry keratoconjunctivitis caused by tear deficiency or excessive evaporation of tears.

Blepharitis is a chronic disorder that causes inflammation of the anterior and posterior edges of the eyelids, involving the skin and related structures (hair and sebaceous glands), mucocutaneous junction, and meibomian glands. It can also affect the conjunctiva, the tear film, and the surface of the cornea in the last stages, and may be associated with a dry eye. Blepharitis is usually classified as anterior or posterior blepharitis, with the anterior one affecting the eyelid region that carries the eyelashes, while the posterior one mainly affects the mouth of the meibomian glands.

Meibomian gland disease is most common in one of three forms: primary meibomite, secondary meibomite, and meibomian seborrhea. Meibomian seborrhea is characterized by excessive secretion of the meibomian glands in the absence of inflammation (hypersecretory disease of the meibomian glands). Primary meibomite, in contrast, is recognized by stagnant and thickened secretion of the meibomian glands (obstructive hypersecretory disease of the meibomian gland). Secondary meibomite, is a localized inflammatory response in which the meibomian glands are secondarily inflamed in a heterogeneous manner, due to anterior blepharitis of the adjacent eyelid.

Weakened corneal sensitivity often occurs after refractive surgery, such as subepithelial laser keratomileusis (SCPL), EPI-SCPL, individual transepithelial non-contact ablation, or other procedures in which the corneal nerves are cut. Weakened corneal sensitivity may also occur after a viral infection caused by HSV-1, HSV-2 and VZV viruses. Patients with weakened corneal sensitivity often complain of a feeling of dry eyes, even if the production and evaporation of tears can be normal, meaning that “dryness” in such patients is actually a form of corneal neuropathy that occurs when the corneal nerves are cut surgically or inflamed after viral infection.

Allergic conjunctivitis is an inflammation of the conjunctiva resulting from hypersensitivity to one or more allergens. It can be acute, intermittent, or chronic. It happens seasonally, that is, only at a certain time of the year, or constantly, that is, chronically throughout the year. Symptoms of seasonal and persistent allergic conjunctivitis, in addition to conjunctival inflammation, include lacrimation, lacrimation, conjunctival vascular distension, pruritus, papillary hyperplasia, chemosis, eyelid edema, and eye discharge. Excretion may form a crust around the eyes after a night's sleep.

Atopic keratoconjunctivitis is a chronic, severe form of allergic conjunctivitis, which often leads to impaired vision. Symptoms include itching, burning sensation, pain, redness, foreign body sensation, light sensitivity, and blurry vision. Often, discharge occurs, especially when waking up after a night's sleep; secretions can be viscous, sticky, and mucous. The inferior conjunctiva is often more affected than the superior conjunctiva. The conjunctiva can vary in the range from pale, edematous and not having features, to having characteristics of the progression of the disease, including papillary hypertrophy, subepithelial fibrosis, arch shortening, trichiasis, eyelid inversion and madarosis. In some patients, the disease progresses to point epithelial erosion, corneal neovascularization, and other manifestations of corneal lesions that can impair vision. Usually there is an overgrowth of goblet cells in the conjunctiva, the formation of a pseudotubular epithelium, and an increased number of degranulating eosinophils and mast cells in the epithelium. The number of CD25 + T lymphocytes, macrophages, dendritic cells (HLA-DR.sup. +, HLA-CD1 +) is significantly increased in its own substance.

Like atopic keratoconjunctivitis, spring keratoconjunctivitis is a severe form of allergic conjunctivitis, but it usually affects the upper conjunctiva more than the lower. It appears in two forms. In palpebral form, there are square, solid, rigid, oblate, closely packed papillae; in the bulbar (marginal) form, the surrounding conjunctiva becomes hypertrophic and grayish. Both forms are often accompanied by mucous secretions. Loss of corneal epithelium may occur, accompanied by pain and photophobia, and plaques may also appear in the central cornea and Trantas point.

EXAMPLES

The invention is further illustrated by the following examples.

Example 1

The inventors prepared the following pharmaceutical formulations:

CMC = carboxymethyl cellulose

HEC = hydroxyethyl cellulose

HPMC = hydroxypropyl methyl cellulose

PVP = polyvinylpyrrolidone

qs = in sufficient quantity

The compositions of table 1 were prepared according to the following procedure:

1. Preparation of concentrated nanosuspensions of Cyclosporin A

but. Cyclosporin A was mixed with an appropriate suspension medium.

The concentration of cyclosporin A in this suspension is in the range of 1-10%.

b. A suspension of Cyclosporin A was crushed using a high pressure homogenizer (Microfluidizer®, manufactured by Microfluidics, Newton, MA), or a ball mill to form a nanosuspension such that d90 <11 μm.

at. The media used to prepare the nanosuspension concentrate are listed in Table 1

2. Preparation of the final product

but. The concentrated nanosuspensions prepared in step 1 were diluted in media suitable for ophthalmic dosing to obtain the final product with the required dose efficiency of cyclosporin A. Media suitable for dilution may contain buffer solutions, stabilizers, gelling agents, and / or a diluent to obtain the final formulations with the desired concentration of CsA.

b. The pharmaceutical compositions of the nanosuspension compositions prepared after dilution of the nanosuspension concentrates are listed in Table 2,

The particle size distribution in various formulations is illustrated in Figures 1, 2 and 3.

Figure 1 illustrates the particle size distribution of Cyclosporin A of Form 2 in nanosuspensions prepared using a high pressure homogenizer, compared with suspensions of pre-pulverization.

Figure 2 compares the particle size distribution of Cyclosporin A of Form 2 in nanosuspensions prepared using various surfactants and stabilizers.

Figure 3 illustrates the particle size distribution of the Composition diluted in a medium containing Na CMC. (Table 2, medium A). It should be noted that no change in particle size is observed in diluted formulations after 2 weeks, compared with the nanosuspension concentrate.

The average particle size of Cyclosporin A Form 3 is illustrated in Figure 4. Cyclosporin A Form 2 forms nanosuspensions (particle size <1 μm), while Form 3 does not.

These experiments show that Form 2 uniformly forms nanosuspensions with a lower particle size than any other crystalline form of CsA. The small particle size of the nanopowders of Form 2 is an advantage over other forms; among other prerequisites, they are expected to exhibit higher bioavailability due to the greater surface area for dissolution, and longer retention on the eye, as well as improved physical stability.

Example 2

The inventors prepared nanosuspensions of Cyclosporin A listed in table 2 below:

PS80 = Polysorbate 80

The inventors tested the above formulations on a New Zealand (NZW) breed of female rabbits (two rabbits in total, one composition in each eye) in a single dose of topical application. The results are summarized in Figures 5-13. They suggest that composition B1 provides a 5-fold lower dose of cyclosporine (0.01% compared with 0.05%), while maintaining a corneal effect comparable to 0.05% Restasis®, and also improves delivery to the bulbar and palpebral conjunctiva.

Claims (15)

1. An eye suspension containing cyclosporin A form 2 and any medium where cyclosporin A form 2 is characterized by the following peaks when scanned by an X-ray diffractometer with Cu Kα, 2θ radiation: 7.5, 8.8, 10.2, 11.3 , 12.7, 13.8, 14.5, 15.6 and 17.5. 2. An ophthalmic suspension according to claim 1, wherein the medium contains at least one surfactant and at least one stabilizer. 3. An ophthalmic suspension according to claim 2, in which cyclosporin is present in a concentration of from about 0.01% to about 10%. 4. An eye suspension according to claim 3, in which the medium contains one, two, three, four, five or six surfactants, each of which is selected from the group consisting of polyoxyethylene (20) sorbitan monooleate, polyethylene glycol 660 hydroxystearate, polyoxyethylene (40) stearate, pluronic F68, polyoxyethylene sorbitan monolaurate, sodium glycocholate. 5. An ophthalmic suspension according to claim 3, in which the medium contains one, two, three, four, five or six surfactants, each of which is selected from the group consisting of polyoxyethylene (20) sorbitan monooleate in concentrations from about 0.1 % to about 5% (weight / volume), polyethylene glycol 660 hydroxystearate in a concentration of from about 0.1% to about 5% (weight / volume), polyoxyethylene (40) stearate in a concentration of from about 0.1% to about 5% (mass / volume), pluronic F68 in a concentration of from about 0.1% to about 5% (mass / volume), polyoxyethylene sorbitan monolaurate in a concentration of from about 0.1% to about 5% (w / v), and sodium glycocholate in a concentration of from about 0.1% to about 5% (w / v). 6. An eye suspension according to claim 4, wherein the medium contains one, two, three or four stabilizers, each of which is selected from the group consisting of hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, polyvinylpyrrolidone and carboxymethyl cellulose. 7. An eye suspension according to claim 5, in which the medium contains one, two, three or four stabilizers, each of which is selected from the group comprising hydroxypropyl cellulose in a concentration of from about 0.1% to about 5% (weight / volume), hydroxypropyl methylcellulose in a concentration of from about 0.1% to about 5% (mass / volume), hydroxyethyl cellulose in a concentration of from about 0.1% to about 5% (mass / volume), and polyvinylpyrrolidone in a concentration of from about 0.1% to about 5% (w / v), and carboxymethyl cellulose in a concentration of from about 0.1% to about 5% (w / v). 8. An eye suspension according to claim 7, wherein the medium further comprises one, two, three, four, five, six, seven, eight or nine ingredients selected from glycerol, mannitol, Pemulen TR-2, sodium citrate dihydrate, potassium chloride , boric acid, sodium borate decahydrate and water. 9. An eye suspension according to claim 7, in which the medium additionally contains one, two, three, four, five, six, seven, eight or nine ingredients selected from glycerol in a concentration of from about 1.0% to about 2.2 %, mannitol at a concentration of about 0.5%, Pemulen TR-2 at a concentration of from about 0.01% to about 0.1%, sodium citrate dihydrate at a concentration of about 0.4%, potassium chloride at a concentration of about 0.14% , boric acid at a concentration of about 0.25%, sodium borate decahydrate at a concentration of about 0.41%, and water. 10. A method of preparing a cyclosporin composition, which includes the steps of mixing cyclosporin A form 2 with a suspension medium; grinding of the suspension, where cyclosporin A form 2 is characterized by the following peaks when scanning with an X-ray diffractometer with radiation Cu Kα, 2θ: 7.5, 8.8, 10.2, 11.3, 12.7, 13.8, 14.5, 15.6 and 17.5. 11. The method according to p. 10, in which the suspension is crushed in the range from about 10 minutes to about 120 minutes. 12. The method according to p. 10, in which the suspension is crushed for about 60 minutes. 13. An ocular suspension containing particles of cyclosporin A form 2; and a medium in which the average particle size (d90) is less than about 10 μm, where cyclosporin A of form 2 is characterized by the following peaks when scanned with a Cu Kα, 2θ X-ray diffractometer: 7.5, 8.8, 10.2, 11, 3, 12.7, 13.8, 14.5, 15.6 and 17.5. 14. An eye suspension according to claim 13, in which the average particle size is less than about 1 μm. 15. A method for treating a condition selected from: dry eye cider, blepharitis, meibomian gland disease, corneal sensitivity disorder, allergic conjunctivitis, atopic keratoconjunctivitis, spring keratoconjunctivitis and pterygium, comprising the step of applying to a patient having such a condition, ocular suspension according to any one of . 1-9, 13 and 14.

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